Table 1.
Strains and plasmids used in the study
| Strains and plasmids | Characteristics | Source |
|---|---|---|
| Brucella abortus | ||
| S2308 | Wild type strain; Smooth phenotype | ATCC |
| ∆pyk | pyk gene deletion mutant strain; Smooth phenotype | This study |
| ∆pyk(Pyk-3 × Flag) | Cmr; complementation strain; ∆pyk carrying the complementation plasmid pBBR-pyk-3 × Flag; Smooth phenotype | This study |
| Escherichia coli | ||
| DH5α | F− φ80lacZ∆M15 ∆(lacZYA-argF)U169 recA1 endA1 hsdR17(r−k, m+k) phoA supE44 thi-1 gyrA96 relA1 λ− | Invitrogen |
| Plasmids | ||
| pBBR1MCS1 | Cmr; Broad-host-range cloning vector | [17] |
| pSC | Ampr; pUC19 plasmid containing SacB gene | [16] |
| pSC-∆pyk | Ampr; pSC plasmid containing the ∆pyk fragment; used to construct deletion strain | This study |
| p3 × Flag-CMV-14 | Ampr; eukaryotic expression plasmid | Sigma-Aldrich |
| pBBR-pyk-3 × Flag | Cmr; pBBR1MCS1 containing the pyk gene flanked by its upstream and downstream regions containing a C-terminal 3 × flag tag. | This study |